Computationally Guided Design of a Soluble, Abundant and Functional AnfH Variant for the Expression of Fe-Only Nitrogenase in Plant Mitochondria.

ABSTRACT: Engineering nitrogenase directly into crops is a long-held aspiration in plant biotechnology. Of the three types, the Fe-only nitrogenase is a promising engineering target, as it has a simpler maturation pathway than the MoFe- and VFe-nitrogenases and does not require any heterometals for its cofactor. However, previously, we have reported that the obligate electron donor of the Fe-only nitrogenase from A. vinelandii, AnfH, is mostly insoluble when expressed in plant mitochondria. Here, we employed computational methods, the Protein Repair One-Stop Shop (PROSS) protein optimisation algorithm and Rosetta energy calculations, to design eight variants of AnfH with improved soluble expression. The amino acid substitutions chosen were predicted to lower the free energy of the native state of th

SUBMITTER: Gregg CM 

PROVIDER: S-EPMC12665077 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature